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An end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics

Next-generation multi-specific antibody therapeutics (MSATs) are engineered to combine several functional activities into one molecule to provide higher efficacy compared to conventional, mono-specific antibody therapeutics. However, highly engineered MSATs frequently display poor yields and less fa...

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Autores principales: Furtmann, Norbert, Schneider, Marion, Spindler, Nadja, Steinmann, Bjoern, Li, Ziyu, Focken, Ingo, Meyer, Joachim, Dimova, Dilyana, Kroll, Katja, Leuschner, Wulf Dirk, Debeaumont, Audrey, Mathieu, Magali, Lange, Christian, Dittrich, Werner, Kruip, Jochen, Schmidt, Thorsten, Birkenfeld, Joerg
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8366542/
https://www.ncbi.nlm.nih.gov/pubmed/34382900
http://dx.doi.org/10.1080/19420862.2021.1955433
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author Furtmann, Norbert
Schneider, Marion
Spindler, Nadja
Steinmann, Bjoern
Li, Ziyu
Focken, Ingo
Meyer, Joachim
Dimova, Dilyana
Kroll, Katja
Leuschner, Wulf Dirk
Debeaumont, Audrey
Mathieu, Magali
Lange, Christian
Dittrich, Werner
Kruip, Jochen
Schmidt, Thorsten
Birkenfeld, Joerg
author_facet Furtmann, Norbert
Schneider, Marion
Spindler, Nadja
Steinmann, Bjoern
Li, Ziyu
Focken, Ingo
Meyer, Joachim
Dimova, Dilyana
Kroll, Katja
Leuschner, Wulf Dirk
Debeaumont, Audrey
Mathieu, Magali
Lange, Christian
Dittrich, Werner
Kruip, Jochen
Schmidt, Thorsten
Birkenfeld, Joerg
author_sort Furtmann, Norbert
collection PubMed
description Next-generation multi-specific antibody therapeutics (MSATs) are engineered to combine several functional activities into one molecule to provide higher efficacy compared to conventional, mono-specific antibody therapeutics. However, highly engineered MSATs frequently display poor yields and less favorable drug-like properties (DLPs), which can adversely affect their development. Systematic screening of a large panel of MSAT variants in very high throughput (HT) is thus critical to identify potent molecule candidates with good yield and DLPs early in the discovery process. Here we report on the establishment of a novel, format-agnostic platform process for the fast generation and multiparametric screening of tens of thousands of MSAT variants. To this end, we have introduced full automation across the entire value chain for MSAT engineering. Specifically, we have automated the in-silico design of very large MSAT panels such that it reflects precisely the wet-lab processes for MSAT DNA library generation. This includes mass saturation mutagenesis or bulk modular cloning technologies while, concomitantly, enabling library deconvolution approaches using HT Sanger DNA sequencing. These DNA workflows are tightly linked to fully automated downstream processes for compartmentalized mammalian cell transfection expression, and screening of multiple parameters. All sub-processes are seamlessly integrated with tailored workflow supporting bioinformatics. As described here, we used this platform to perform multifactor optimization of a next-generation bispecific, cross-over dual variable domain-Ig (CODV-Ig). Screening of more than 25,000 individual protein variants in mono- and bispecific format led to the identification of CODV-Ig variants with over 1,000-fold increased potency and significantly optimized production titers, demonstrating the power and versatility of the platform.
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spelling pubmed-83665422021-08-17 An end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics Furtmann, Norbert Schneider, Marion Spindler, Nadja Steinmann, Bjoern Li, Ziyu Focken, Ingo Meyer, Joachim Dimova, Dilyana Kroll, Katja Leuschner, Wulf Dirk Debeaumont, Audrey Mathieu, Magali Lange, Christian Dittrich, Werner Kruip, Jochen Schmidt, Thorsten Birkenfeld, Joerg MAbs Report Next-generation multi-specific antibody therapeutics (MSATs) are engineered to combine several functional activities into one molecule to provide higher efficacy compared to conventional, mono-specific antibody therapeutics. However, highly engineered MSATs frequently display poor yields and less favorable drug-like properties (DLPs), which can adversely affect their development. Systematic screening of a large panel of MSAT variants in very high throughput (HT) is thus critical to identify potent molecule candidates with good yield and DLPs early in the discovery process. Here we report on the establishment of a novel, format-agnostic platform process for the fast generation and multiparametric screening of tens of thousands of MSAT variants. To this end, we have introduced full automation across the entire value chain for MSAT engineering. Specifically, we have automated the in-silico design of very large MSAT panels such that it reflects precisely the wet-lab processes for MSAT DNA library generation. This includes mass saturation mutagenesis or bulk modular cloning technologies while, concomitantly, enabling library deconvolution approaches using HT Sanger DNA sequencing. These DNA workflows are tightly linked to fully automated downstream processes for compartmentalized mammalian cell transfection expression, and screening of multiple parameters. All sub-processes are seamlessly integrated with tailored workflow supporting bioinformatics. As described here, we used this platform to perform multifactor optimization of a next-generation bispecific, cross-over dual variable domain-Ig (CODV-Ig). Screening of more than 25,000 individual protein variants in mono- and bispecific format led to the identification of CODV-Ig variants with over 1,000-fold increased potency and significantly optimized production titers, demonstrating the power and versatility of the platform. Taylor & Francis 2021-08-12 /pmc/articles/PMC8366542/ /pubmed/34382900 http://dx.doi.org/10.1080/19420862.2021.1955433 Text en © 2021 The Author(s). Published with license by Taylor & Francis Group, LLC. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) ), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Report
Furtmann, Norbert
Schneider, Marion
Spindler, Nadja
Steinmann, Bjoern
Li, Ziyu
Focken, Ingo
Meyer, Joachim
Dimova, Dilyana
Kroll, Katja
Leuschner, Wulf Dirk
Debeaumont, Audrey
Mathieu, Magali
Lange, Christian
Dittrich, Werner
Kruip, Jochen
Schmidt, Thorsten
Birkenfeld, Joerg
An end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics
title An end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics
title_full An end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics
title_fullStr An end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics
title_full_unstemmed An end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics
title_short An end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics
title_sort end-to-end automated platform process for high-throughput engineering of next-generation multi-specific antibody therapeutics
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8366542/
https://www.ncbi.nlm.nih.gov/pubmed/34382900
http://dx.doi.org/10.1080/19420862.2021.1955433
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